Fig. 4.

Immunoprecipitation of modified pDsRed2-C1-1NLS and pDsRed2-C1-S100A10-1NLS. Immunoprecipitation was performed using COS1 and COS7 cell lines. pDsRed2-C1-wt (Clontech), pDsRed2-C1-1NLS, pDsRed2-C1-S100A10-wt, and pDsRed2-C1-S100A10-1NLS vectors were transfected into each cell line for 24 h. Cells were lysed after transfection and immunoprecipitation was performed in the lysate using DsRed, karyopherin α2/β2 antibodies, and protein A/G plus-agarose (Santa Cruz). Samples that transfected with pDsRed2-C1-wt (Clontech), pDsRed2-C1-1NLS, pDsRed2-C1-S100A10-wt, pDsRed2-C1-S100A10-1NLS were identified using antibodies that detected karyopherin α2/β2 protein. Samples evaluated for karyopherin α2/β2 were confirmed by western blotting using antibodies for DsRed protein. Anti-β-actin (C4) antibody (Santa Cruz) was used to detect a reference protein in whole cell lysates prior to immunoprecipitation